Site-specific Antibody Service


  • PTMs (Post-Translational Modifications) are an important consideration in research. They contribute to a variety of biological activities and affect protein structures and cellular functions. Protein phosphorylation, methylation, and acetylation are the most common types of modifications of the more than 200 different attached modifications known to exist in vivo.
  • These site-specific antibodies have a number of potential applications, including: cell signaling, protein structure, function, development, maturation, localization, disease diagnostics, activity, etc. Over the years, ABclonal has developed thousands of custom site-specific antibodies and has become a certifiable expert in the custom site-specific antibody services.
  • All custom modification antibody productions will undergo double affinity purification with modified and unmodified peptides.

PTMs in the Cell

The major types of post-translational modifications (Jensen O. N. 2006)

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Phosphorylation Acetylation Methylation Ubiquitination
Citrullination Succinylation Alkylation Isoprenylation
Prenylation Sumoylation Tyrosine Sulfation Tyrosine Nitration
Glycosylation Biotinylation Polyglutamylation Other PTMs

Package


  • Peptide design and synthesis
  • Customer-supplied peptide
  • Peptide-KLH conjugation
  • Immunization of 2 rabbits
  • ELISA titer test
  • Antigen affinity column purification

Note:

1. If the antigen is provided by the customer: modified peptide (<15 aa >90% purity, 24 mg) and unmodified control (<15 aa >90% purity, 11 mg) are needed.

Accord to our years of experience, the success rate of the modified antibody preparation is lower than the normal one. To ensure the probability of obtaining a useful antibody after immunization, we recommend to immunize 3-4 rabbits.

Modified synthetic peptide usually has a chain of 14 amino acids. Approximately 10 mg are used in modified epitope affinity purification, 10 mg are used in antibody production, and 4 mg are used in QC.

Approximately 10 mg of non-modified peptide (about 14 amino acids) are used in affinity purification.

To obtain a more specific modified antibody, sometimes we used shortened modified peptides to purify.

Professional Antigen Design (Free)MORE


Proper design of a suitable antigen is the most critical step in getting a useful antibody.

High-quality antigen preparationMORE


Through professional analysis, we choose the best region to synthesize peptide coupling KLH as the antigen to immune animals.

Each synthesized peptide goes through HPLC and MS tests to ultimately obtain a useful antibody.

Other Modifications


ABclonal offers DNA methylation, RNA methylation, pan-acetylation, non-histone modification and other custom specific modified antibodies that are developed to meet your research needs.

Use Nipponbare callus (same quantity, about 3 g) as sample material of ChIP experiment, 02g and 03g as target gene. Conduct IP experiment use anti-H3K27me3 (ABclonal) and anti-H3K27me3 (Company M).

Conclusion:
The efficiency of ABclonal anti-H3K27me3 immunoprecipitation is 10 times that of Company M anti-H3K27me3.

High Throughput Detection


Obtaining a useful site-specific antibody is not easy. Most manufacturers cannot achieve this because of antibody validation. In addition to manufacturing useful site-specific antibodies, we dedicate ourselves to be the best validation platform in the world. ABclonal has built advanced testing platforms for Western Blot, immunohistochemistry, and immunofluorescence-based assays, supplemented by immunoprecipitation, RIP, ChIP, ChIP-seq, and shRNA knockout validation. We take every measure to provide the most accurate and reliable data for each of our customers.

ABclonal's standard protocol for site-specific antibody is optimized to produce antiserum in the shortest possible time. It is the most affordable option and a popular choice among researchers.

The protocol uses two New Zealand White rabbits that are Specific-Pathogen-Free. Injections are subcutaneous (SQ) as emulsions in Complete Freund's Adjuvant (CFA) or Incomplete Freund's Adjuvant (IFA). We guarantee that the same antigen will be injected into both animals.

Procedure Protocol Day Description
Control Serum Collection Day 0 Pre-immune bleed (5 mL per rabbit)
Primary Injection Day 1 Immunize with 0.50 mg of antigen in CFA, 10 SQ sites
1st Booster Day 14 Boost with 0.25 mg of antigen in IFA, 4 SQ sites
2nd Booster Day 28 Boost with 0.25 mg of antigen in IFA, 4 SQ sites
Serum Collection Day 35 Bleed (-25 mL per rabbit)
3rd Booster Day 42 Boost with 0.25 mg of antigen in IFA, 4 SQ sites
Serum Collection Day 56, 58 Two bleeds (total of -50 mL per rabbit)
ELISA Test Day 60 ELISA titration (results available online)
Verify disposition of rabbits
Decide to continue or terminate
Purification Optional 6-10 mg of purified antibodies
Note: After Day 35, the scheduled days in this protocol are approximations and are dependent on the titer test.
Quote
  1. For quotation requests and questions, you may contact us by email, phone, or via our online quotation system.
  2. Orders can be placed by phone, email, fax, or online with a formal Purchase Order or credit card.
  3. Please send your samples to ABclonal, 86 Cummings Park Dr, Woburn, MA 01801, United States.
  4. Our customer service representatives are available 24/7.